GSM8084141: ATAC-Rreb1KO_TGFb_Rep2; Mus musculus; ATAC-seq
samples containing 5x104 cells were washed twice with cold ATAC-RSB (10 mM Tris-HCl pH 7.5, 10 mM NaCl, 3 mM MgCl2) and resuspended with ice-cold ATAC-lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Nonidet P-40, 0.1% Tween-20, 0.01% digitonin), followed by incubation on ice for 3 min. After washing the cells with cold ATAC-wash buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20), cells were resuspended with 50 μL of transposition mix (10 mM Tris-HCl pH 7.5, 5 mM MgCl2, 10% dimethyl formamide, 100 nM Nextera TDE1 (Illumina, Cat #: FC-121–1030), 0.01% Digitonin, 0.1% Tween-20) and incubated at 37ºC for 30 min in a thermomixer with 1000 rpm mixing. DNA was purified using a QIAquick PCR Purification Kit (Qiagen). ATAC-seq libraries were prepared using the NEBNext High-Fidelity 2× PCR Master Mix (NEB, Cat #: M0541). Libraries were purified by double-sided size selection with AMPure XP beads (Beckman Coulter) and assessed using a Bioanalyzer High-Sensitivity DNA Analysis kit (Agilent, Cat #: 5067-4626).